Compositions and methods for treating wilson's disease

Abstract

Provided herein are nucleic acid molecules and vectors comprising variant copper transporting ATPase 2 (ATP7B) nucleic acid sequences. Such sequences have been optimized for expression in mammalian cells, liver cells, and/or from an adeno associated viral vector (AAV), including truncated and/or codon optimized variants. Also provided are viral vectors comprising such ATP7B variant nucleic acid sequences, and methods of use thereof for treating disorders associated with an ATP7B deficiency, such as Wilsons disease.

What is claimed is: 
     
         1 . A nucleic acid molecule comprising a nucleotide sequence having (i) at least 80% sequence identity to SEQ ID NO: 3 or 4, or (ii) at least 92% sequence identity to SEQ ID NO: 5 or 6, wherein said nucleic acid molecule encodes a functional ATP7B protein. 
     
     
         2 . The nucleic acid molecule of  claim 1 , comprising a nucleotide sequence having at least 85% sequence identity to SEQ ID NO: 3 or 4. 
     
     
         3 . The nucleic acid molecule of  claim 1 , comprising a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 3 or 4. 
     
     
         4 . The nucleic acid molecule of  claim 1 , comprising a nucleotide sequence having at least 95% sequence identity to any one of SEQ ID NOs: 3-6. 
     
     
         5 . The nucleic acid molecule of  claim 1 , comprising a nucleotide sequence having any one of SEQ ID NOs: 3-6. 
     
     
         6 . The nucleic acid molecule of  claim 1 , wherein the nucleic acid molecule encodes a functional fragment of ATP7B. 
     
     
         7 . The nucleic acid molecule of  claim 1 , wherein the nucleic acid molecule encodes an ATP7B protein having SEQ ID NO: 2. 
     
     
         8 . The nucleic acid molecule of any one of  claims 1 - 7 , wherein the level of expression of the functional ATP7B protein from the nucleic acid molecule in the liver is at least 3-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         9 . The nucleic acid molecule of  claim 8 , wherein the level of expression of the functional ATP7B protein from the nucleic acid molecule in the liver is at least 5-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         10 . The nucleic acid molecule of  claim 9 , wherein the level of expression of the functional ATP7B protein from the nucleic acid molecule in the liver is at least 7-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         11 . The nucleic acid molecule of any one of  claims 1 - 10 , further comprising a regulatory element. 
     
     
         12 . The nucleic acid molecule of  claim 11 , wherein the regulatory element comprises a sequence having at least 85% sequence identity to any one of SEQ ID NOs: 19-43. 
     
     
         13 . The nucleic acid molecule of  claim 12 , wherein the regulatory element comprises a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 19-43. 
     
     
         14 . The nucleic acid molecule of  claim 13 , wherein the regulatory element comprises a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 19-43. 
     
     
         15 . The nucleic acid molecule of  claim 14 , wherein the regulatory element comprises a sequence having any one of SEQ ID NOs: 19-43. 
     
     
         16 . The nucleic acid molecule of any one of  claims 11 - 15 , wherein the regulatory element comprises a promoter sequence. 
     
     
         17 . The nucleic acid molecule of  claim 16 , wherein the promoter sequence produces at least 10-fold greater expression in a mammalian cell relative to the CMV promoter. 
     
     
         18 . The nucleic acid molecule of  claim 16 , wherein the promoter sequence produces at least 50-fold greater expression in a mammalian cell relative to the CMV promoter. 
     
     
         19 . The nucleic acid molecule of any one of  claims 11 - 18 , wherein the regulatory element further comprises an enhancer sequence. 
     
     
         20 . The nucleic acid molecule of any one of  claims 11 - 19 , wherein the regulatory element has less than 150 bp. 
     
     
         21 . The nucleic acid molecule of any one of  claims 11 - 19 , wherein the regulatory element has less than 120 bp. 
     
     
         22 . The nucleic acid molecule of any one of  claims 11 - 19 , wherein the regulatory element has less than 105 bp. 
     
     
         23 . The nucleic acid molecule of any one of  claims 1 - 22 , further comprising a 5′ ITR and 3′ ITR sequences of a virus. 
     
     
         24 . The nucleic acid molecule of  claim 23 , wherein the 5′ ITR and 3′ ITR sequences are adeno-associated virus (AAV) sequences. 
     
     
         25 . The nucleic acid molecule of  claim 24 , wherein the AAV is an AAV2, AAV5, AAV8 or AAV9. 
     
     
         26 . An expression vector comprising the nucleic acid molecule of any one of  claims 1 - 25 . 
     
     
         27 . The expression vector of  claim 26 , wherein the expression vector is a viral vector. 
     
     
         28 . The expression vector of  claim 27 , wherein the viral vector is an AAV vector. 
     
     
         29 . A viral particle comprising the nucleic acid molecule of any one of  claims 1 - 25  or the expression vector of any one of  claims 26 - 28 . 
     
     
         30 . The viral particle of  claim 29 , wherein the viral particle comprises capsid proteins of an AAV. 
     
     
         31 . The viral particle of  claim 30 , wherein the AAV is an AAV5, AAV8 or AAV9. 
     
     
         32 . A host cell comprising the nucleic acid molecule of any one of  claims 1 - 25  or the expression vector of any one of  claims 26 - 28 . 
     
     
         33 . A pharmaceutical composition comprising the nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , or the viral particle of any one of  claims 29 - 31 , and one or more pharmaceutically acceptable excipients. 
     
     
         34 . A method for increasing expression of functional ATP7B protein in a subject comprising administering to said subject the nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , the viral particle of any one of  claims 29 - 31 , or the pharmaceutical composition of  claim 33 . 
     
     
         35 . A method for treating a disorder associated with an ATP7B deficiency comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , the viral particle of any one of  claims 29 - 31 , or the pharmaceutical composition of  claim 33 . 
     
     
         36 . A method for treating Wilson's disease comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , the viral particle of any one of  claims 29 - 31 , or the pharmaceutical composition of  claim 33 . 
     
     
         37 . The nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , the viral particle of any one of  claims 29 - 31 , or the pharmaceutical composition of  claim 33  for use in increasing the expression of functional ATP7B in a subject. 
     
     
         38 . The nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , the viral particle of any one of  claims 29 - 31 , or the pharmaceutical composition of  claim 33  for use in increasing treating a disorder associated with an ATP7B deficiency. 
     
     
         39 . The nucleic acid molecule of any one of  claims 1 - 25 , the expression vector of any one of  claims 26 - 28 , the viral particle of any one of  claims 29 - 31 , or the pharmaceutical composition of  claim 33  for use in treating Wilson's disease. 
     
     
         40 . A process of producing a viral particle according to any of  claims 29 - 31 , comprising: (i) culturing a host cell according to  claim 32  in a culture medium, and (ii) harvesting the viral particles from the cell culture supernatant or the host cells. 
     
     
         41 . The process of  claim 40 , wherein the host cell further comprises (a) a nucleic acid molecule encoding AAV rep and/or cap genes, and/or (b) a nucleic acid molecule comprising viral helper genes. 
     
     
         42 . Use of the nucleic acid molecule of any of  claims 1 - 25  or the expression vector of any one of  claims 26 - 28  for the production of viral particles. 
     
     
         43 . A nucleic acid molecule comprising:
 (i) a regulatory element comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 19-43, and   (ii) a variant nucleotide sequence encoding a functional ATP7B protein, wherein the variant nucleotide sequence has been optimized for expression in liver from a viral vector.   
     
     
         44 . The nucleic acid molecule of  claim 43 , wherein the regulatory element comprises a sequence having at least 85% sequence identity to any one of SEQ ID NOs: 19-43. 
     
     
         45 . The nucleic acid molecule of  claim 44 , wherein the regulatory element comprises a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 19-43. 
     
     
         46 . The nucleic acid molecule of  claim 45 , wherein the regulatory element comprises a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 19-43. 
     
     
         47 . The nucleic acid molecule of  claim 46 , wherein the regulatory element comprises a sequence having any one of SEQ ID NOs: 19-43. 
     
     
         48 . The nucleic acid molecule of any one of  claims 43 - 47 , wherein the variant nucleotide sequence has been codon optimized for expression in the liver. 
     
     
         49 . The nucleic acid molecule of any one of  claims 43 - 48 , wherein the variant nucleotide sequence has been optimized for expression from an AAV vector. 
     
     
         50 . The nucleic acid molecule of any one of  claims 43 - 49 , wherein the variant nucleotide sequence has least 80% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         51 . The nucleic acid molecule of  claim 50 , wherein the variant nucleotide sequence has at least 85% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         52 . The nucleic acid molecule of  claim 51 , wherein the variant nucleotide sequence has at least 90% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         53 . The nucleic acid molecule of  claim 52 , wherein the variant nucleotide sequence has at least 95% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         54 . The nucleic acid molecule of  claim 53 , wherein the variant nucleotide sequence comprises any one of SEQ ID NOs: 3-18. 
     
     
         55 . The nucleic acid molecule of any one of  claims 43 - 54 , wherein the regulatory element is a promoter sequence. 
     
     
         56 . The nucleic acid molecule of any one of  claims 43 - 55 , further comprising an enhancer sequence. 
     
     
         57 . The nucleic acid molecule of any one of  claims 43 - 56 , wherein the regulatory element has less than 150 bp. 
     
     
         58 . The nucleic acid molecule of any one of  claims 43 - 56 , wherein the regulatory element has less than 120 bp. 
     
     
         59 . The nucleic acid molecule of any one of  claims 43 - 56 , wherein the regulatory element has less than 105 bp. 
     
     
         60 . The nucleic acid molecule of any one of  claims 43 - 59 , wherein the regulatory element produces at least 10-fold greater expression in a mammalian cell relative to the CMV promoter. 
     
     
         61 . The nucleic acid molecule of any one of  claims 43 - 60 , wherein the regulatory element produces at least 50-fold greater expression in a mammalian cell relative to the CMV promoter. 
     
     
         62 . The nucleic acid molecule of any one of  claims 43 - 61 , wherein the nucleic acid molecule encodes a functional fragment of ATP7B. 
     
     
         63 . The nucleic acid molecule of any one of  claims 43 - 61 , wherein the nucleic acid molecule encodes an ATP7B protein having SEQ ID NO: 2. 
     
     
         64 . The nucleic acid molecule of any one of  claims 43 - 63 , wherein the level of expression of the functional ATP7B protein from the variant nucleotide sequence in the liver is at least 5-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         65 . The nucleic acid molecule of any one of  claims 43 - 63 , wherein the level of expression of the functional ATP7B protein from the variant nucleotide sequence in the liver is at least 7-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         66 . The nucleic acid molecule of any one of  claims 43 - 65 , further comprising a 5′ ITR and 3′ ITR sequences of a virus. 
     
     
         67 . The nucleic acid molecule of  claim 66 , wherein the 5′ ITR and 3′ ITR sequences are adeno-associated virus (AAV) sequences. 
     
     
         68 . The nucleic acid molecule of  claim 67 , wherein the AAV is an AAV2, AAV5, AAV8 or AAV9. 
     
     
         69 . An expression vector comprising the nucleic acid molecule of any one of  claims 43 - 68 . 
     
     
         70 . The expression vector of  claim 69 , wherein the expression vector is a viral vector. 
     
     
         71 . The expression vector of  claim 70 , wherein the viral vector is an AAV vector. 
     
     
         72 . A viral particle comprising the nucleic acid molecule of any one of  claims 43 - 68  or the expression vector of any one of  claims 69 - 71 . 
     
     
         73 . The viral particle of  claim 72 , wherein the viral particle comprises capsid proteins of an AAV. 
     
     
         74 . The viral particle of  claim 73 , wherein the AAV is an AAV5, AAV8 or AAV9. 
     
     
         75 . A host cell comprising the nucleic acid molecule of any one of  claims 43 - 68  or the expression vector of any one of  claims 69 - 71 . 
     
     
         76 . A pharmaceutical composition comprising the nucleic acid molecule of any one of  claims 43 - 68 , the expression vector of any one of  claims 69 - 71 , or the viral particle of any one of  claims 72 - 74 , and one or more pharmaceutically acceptable excipients. 
     
     
         77 . A method for increasing expression of functional ATP7B protein in a subject comprising administering to said subject the nucleic acid molecule of any one of  claims 43 - 68 , the expression vector of any one of  claims 68 - 70 , the viral particle of any one of  claims 72 - 74 , or the pharmaceutical composition of  claim 76 . 
     
     
         78 . A method for treating a disorder associated with an ATP7B deficiency comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 43 - 68 , the expression vector of any one of  claims 69 - 71 , the viral particle of any one of  claims 72 - 74 , or the pharmaceutical composition of  claim 76 . 
     
     
         79 . A method for treating Wilson's disease comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 43 - 68 , the expression vector of any one of  claims 69 - 71 , the viral particle of any one of  claims 72 - 74 , or the pharmaceutical composition of  claim 76 . 
     
     
         80 . The nucleic acid molecule of any one of  claims 43 - 68 , the expression vector of any one of  claims 69 - 71 , the viral particle of any one of  claims 72 - 74 , or the pharmaceutical composition of  claim 76  for use in increasing the expression of functional ATP7B in a subject. 
     
     
         81 . The nucleic acid molecule of any one of  claims 41 - 68 , the expression vector of any one of  claims 69 - 71 , the viral particle of any one of  claims 72 - 74 , or the pharmaceutical composition of  claim 76  for use in increasing treating a disorder associated with an ATP7B deficiency. 
     
     
         82 . The nucleic acid molecule of any one of  claims 43 - 68 , the expression vector of any one of  claims 69 - 71 , the viral particle of any one of  claims 72 - 74 , or the pharmaceutical composition of  claim 76  for use in treating Wilson's disease. 
     
     
         83 . A process of producing a viral particle according to any one of  claims 72 - 74 , comprising: (i) culturing a host cell according to  claim 75  in a culture medium, and (ii) harvesting the viral particles from the cell culture supernatant or the host cells. 
     
     
         84 . The process of  claim 83 , wherein the host cell further comprises (a) a nucleic acid molecule encoding AAV rep and/or cap genes, and/or (b) a nucleic acid molecule comprising viral helper genes. 
     
     
         85 . Use of the nucleic acid molecule of any one of  claims 43 - 68  or the expression vector of any one of  claims 69 - 71  for the production of viral particles. 
     
     
         86 . A nucleic acid molecule comprising:
 (i) a regulatory element comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 66-68, and   (ii) a nucleotide sequence encoding a therapeutic transgene, wherein the therapeutic transgene is operably linked to the regulatory element.   
     
     
         87 . The nucleic acid molecule of  claim 86 , wherein the regulatory element comprises a sequence having at least 85% sequence identity to any one of SEQ ID NOs: 66-68. 
     
     
         88 . The nucleic acid molecule of  claim 87 , wherein the regulatory element comprises a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 66-68. 
     
     
         89 . The nucleic acid molecule of  claim 88 , wherein the regulatory element comprises a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 66-68. 
     
     
         90 . The nucleic acid molecule of  claim 89 , wherein the regulatory element comprises a sequence having any one of SEQ ID NOs: 66-68. 
     
     
         91 . The nucleic acid molecule of  claim 90 , wherein the regulatory element comprises SEQ ID NO: 67. 
     
     
         92 . The nucleic acid molecule of any one of  claims 86 - 91 , wherein the therapeutic transgene encodes any one of ATP7A, ATP7B, ATP8B1, ABCB4, ABCB11, CDKL5, CNTNAP2, ZEB2, Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI or Factor XII, or a variant or functional fragment thereof. 
     
     
         93 . The nucleic acid molecule of  claim 92 , wherein the therapeutic transgene encodes Factor VIII or a variant or functional fragment thereof. 
     
     
         94 . The nucleic acid molecule of  claim 90 , wherein the therapeutic transgene encodes ATP7B or a variant or functional fragment thereof. 
     
     
         95 . The nucleic acid molecule of any one of  claims 86 - 94 , wherein the therapeutic transgene comprises a variant nucleotide sequence that has been codon optimized for expression in the liver. 
     
     
         96 . The nucleic acid molecule of any one of  claims 86 - 95 , wherein the therapeutic transgene comprises a variant nucleotide sequence has been optimized for expression from an AAV vector. 
     
     
         97 . The nucleic acid molecule of any one of  claims 86 - 96 , wherein the therapeutic transgene comprises a variant nucleotide sequence having least 80% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         98 . The nucleic acid molecule of  claim 97 , wherein the therapeutic transgene sequence comprises a variant nucleotide sequence having at least 85% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         99 . The nucleic acid molecule of  claim 98 , wherein the therapeutic transgene sequence comprises a variant nucleotide sequence having at least 90% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         100 . The nucleic acid molecule of  claim 99 , wherein the therapeutic transgene sequence comprises a variant nucleotide sequence having at least 95% sequence identity to any one of SEQ ID NOs: 3-18. 
     
     
         101 . The nucleic acid molecule of  claim 100 , wherein the therapeutic transgene sequence comprises any one of SEQ ID NOs: 3-18. 
     
     
         102 . The nucleic acid molecule of  claim 101 , wherein the therapeutic transgene sequence comprises SEQ ID NO: 5. 
     
     
         103 . The nucleic acid molecule of any one of  claims 86 - 102 , wherein the regulatory element is a promoter sequence. 
     
     
         104 . The nucleic acid molecule of any one of  claims 86 - 103 , further comprising an enhancer sequence. 
     
     
         105 . The nucleic acid molecule of any one of  claims 86 - 104 , wherein the regulatory element has less than 150 bp. 
     
     
         106 . The nucleic acid molecule of any one of  claims 86 - 104 , wherein the regulatory element has less than 120 bp. 
     
     
         107 . The nucleic acid molecule of any one of  claims 86 - 104 , wherein the regulatory element has less than 105 bp. 
     
     
         108 . The nucleic acid molecule of any one of  claims 86 - 107 , wherein the regulatory element produces at least 10-fold greater expression in a mammalian cell relative to the CMV promoter. 
     
     
         109 . The nucleic acid molecule of any one of  claims 86 - 108 , wherein the regulatory element produces at least 50-fold greater expression in a mammalian cell relative to the CMV promoter. 
     
     
         110 . The nucleic acid molecule of any one of  claims 86 - 109 , wherein the therapeutic transgene encodes a functional fragment of ATP7B. 
     
     
         111 . The nucleic acid molecule of any one of  claims 86 - 109 , wherein the therapeutic transgene encodes an ATP7B protein having SEQ ID NO: 2. 
     
     
         112 . The nucleic acid molecule of any one of  claims 86 - 111 , wherein the level of expression of the functional ATP7B protein from the therapeutic transgene sequence in the liver is at least 5-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         113 . The nucleic acid molecule of  claim 112 , wherein the level of expression of the functional ATP7B protein from the therapeutic transgene sequence in the liver is at least 7-fold greater relative to the level of expression of ATP7B protein in the liver from a nucleic acid molecule comprising SEQ ID NO: 1. 
     
     
         114 . The nucleic acid molecule of any one of  claims 86 - 113 , further comprising a 5′ ITR and 3′ ITR sequences of a virus. 
     
     
         115 . The nucleic acid molecule of  claim 114 , wherein the 5′ ITR and 3′ ITR sequences are adeno-associated virus (AAV) sequences. 
     
     
         116 . The nucleic acid molecule of  claim 115 , wherein the AAV is an AAV2, AAV5, AAV8 or AAV9. 
     
     
         117 . An expression vector comprising the nucleic acid molecule of any one of  claims 86 - 116 . 
     
     
         118 . The expression vector of  claim 117 , wherein the expression vector is a viral vector. 
     
     
         119 . The expression vector of  claim 118 , wherein the viral vector is an AAV vector. 
     
     
         120 . A viral particle comprising the nucleic acid molecule of any one of  claims 86 - 116  or the expression vector of any one of  claims 117 - 119 . 
     
     
         121 . The viral particle of  claim 120 , wherein the viral particle comprises capsid proteins of an AAV. 
     
     
         122 . The viral particle of  claim 121 , wherein the AAV is an AAV5, AAV8 or AAV9. 
     
     
         123 . A host cell comprising the nucleic acid molecule of any one of  claims 86 - 116  or the expression vector of any one of  claims 117 - 119 . 
     
     
         124 . A pharmaceutical composition comprising the nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , or the viral particle of any one of  claims 120 - 122 , and one or more pharmaceutically acceptable excipients. 
     
     
         125 . A method for increasing expression of functional ATP7B protein in a subject comprising administering to said subject the nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , the viral particle of any one of  claims 120 - 122 , or the pharmaceutical composition of  claim 124 . 
     
     
         126 . A method for treating a disorder associated with an ATP7B deficiency comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , the viral particle of any one of  claims 120 - 122 , or the pharmaceutical composition of  claim 124 . 
     
     
         127 . A method for treating Wilson's disease comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , the viral particle of any one of  claims 120 - 122 , or the pharmaceutical composition of  claim 124 . 
     
     
         128 . The nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , the viral particle of any one of  claims 120 - 122 , or the pharmaceutical composition of  claim 124  for use in increasing the expression of functional ATP7B in a subject. 
     
     
         129 . The nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , the viral particle of any one of  claims 120 - 122 , or the pharmaceutical composition of  claim 124  for use in increasing treating a disorder associated with an ATP7B deficiency. 
     
     
         130 . The nucleic acid molecule of any one of  claims 86 - 116 , the expression vector of any one of  claims 117 - 119 , the viral particle of any one of  claims 120 - 122 , or the pharmaceutical composition of  claim 124  for use in treating Wilson's disease. 
     
     
         131 . A process of producing a viral particle according to any one of  claims 120 - 122 , comprising: (i) culturing a host cell according to  claim 123  in a culture medium, and (ii) harvesting the viral particles from the cell culture supernatant or the host cells. 
     
     
         132 . The process of  claim 131 , wherein the host cell further comprises (a) a nucleic acid molecule encoding AAV rep and/or cap genes, and/or (b) a nucleic acid molecule comprising viral helper genes. 
     
     
         133 . Use of the nucleic acid molecule of any one of  claims 86 - 116  or the expression vector of any one of  claims 117 - 119  for the production of viral particles. 
     
     
         134 . A nucleic acid molecule comprising:
 (i) a regulatory element comprising a sequence having at least 80% sequence identity to SEQ ID NO: 24, and   (ii) a variant nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 5, wherein said variant nucleotide sequence encodes a functional ATP7B protein.   
     
     
         135 . The nucleic acid molecule of  claim 134 , wherein the regulatory element comprises a sequence having at least 85% sequence identity to SEQ ID NO: 24. 
     
     
         136 . The nucleic acid molecule of  claim 135 , wherein the regulatory element comprises a sequence having at least 90% sequence identity to SEQ ID NO: 24. 
     
     
         137 . The nucleic acid molecule of  claim 136 , wherein the regulatory element comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 24. 
     
     
         138 . The nucleic acid molecule of  claim 137 , wherein the regulatory element comprises a sequence having SEQ ID NO: 24. 
     
     
         139 . The nucleic acid molecule of any one of  claims 134 - 138 , wherein the variant nucleotide sequence comprises a sequence having at least 85% sequence identity to SEQ ID NO: 5. 
     
     
         140 . The nucleic acid molecule of  claim 139 , wherein the variant nucleotide sequence comprises a sequence having at least 90% sequence identity to SEQ ID NO: 5. 
     
     
         141 . The nucleic acid molecule of  claim 140 , wherein the variant nucleotide sequence comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 5. 
     
     
         142 . The nucleic acid molecule of  claim 141 , wherein the variant nucleotide sequence comprises a sequence having SEQ ID NO: 5. 
     
     
         143 . A nucleic acid molecule comprising:
 (i) a regulatory element comprising a sequence having at least 80% sequence identity to SEQ ID NO: 67, and   (ii) a variant nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 5, wherein said variant nucleotide sequence encodes a functional ATP7B protein.   
     
     
         144 . The nucleic acid molecule of  claim 143 , wherein the regulatory element comprises a sequence having at least 85% sequence identity to SEQ ID NO: 67. 
     
     
         145 . The nucleic acid molecule of  claim 144 , wherein the regulatory element comprises a sequence having at least 90% sequence identity to SEQ ID NO: 67. 
     
     
         146 . The nucleic acid molecule of  claim 145 , wherein the regulatory element comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 67. 
     
     
         147 . The nucleic acid molecule of  claim 146 , wherein the regulatory element comprises a sequence having SEQ ID NO: 67. 
     
     
         148 . The nucleic acid molecule of any one of  claims 143 - 147 , wherein the variant nucleotide sequence comprises a sequence having at least 85% sequence identity to SEQ ID NO: 5. 
     
     
         149 . The nucleic acid molecule of  claim 148 , wherein the variant nucleotide sequence comprises a sequence having at least 90% sequence identity to SEQ ID NO: 5. 
     
     
         150 . The nucleic acid molecule of  claim 149 , wherein the variant nucleotide sequence comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 5. 
     
     
         151 . The nucleic acid molecule of  claim 150 , wherein the variant nucleotide sequence comprises a sequence having SEQ ID NO: 5. 
     
     
         152 . The nucleic acid molecule of any one of  claims 134 - 151 , wherein the variant nucleotide sequence encodes a functional fragment of ATP7B. 
     
     
         153 . The nucleic acid molecule of any one of  claims 134 - 152 , wherein the variant nucleotide sequence encodes an ATP7B protein having SEQ ID NO: 2. 
     
     
         154 . The nucleic acid molecule of any one of  claims 134 - 153 , further comprising a 5′ ITR and 3′ ITR sequences of a virus. 
     
     
         155 . The nucleic acid molecule of  claim 154 , wherein the 5′ ITR and 3′ ITR sequences are adeno-associated virus (AAV) sequences. 
     
     
         156 . The nucleic acid molecule of  claim 155 , wherein the AAV is an AAV2, AAV5, AAV8 or AAV9. 
     
     
         157 . An expression vector comprising the nucleic acid molecule of any one of  claims 134 - 156 . 
     
     
         158 . The expression vector of  claim 157 , wherein the expression vector is a viral vector. 
     
     
         159 . The expression vector of  claim 158 , wherein the viral vector is an AAV vector. 
     
     
         160 . A viral particle comprising the nucleic acid molecule of any one of  claims 134 - 156  or the expression vector of any one of  claims 157 - 159 . 
     
     
         161 . The viral particle of  claim 160 , wherein the viral particle comprises capsid proteins of an AAV. 
     
     
         162 . The viral particle of  claim 161 , wherein the AAV is an AAV5, AAV8 or AAV9. 
     
     
         163 . A host cell comprising the nucleic acid molecule of any one of  claims 134 - 156  or the expression vector of any one of  claims 157 - 159 . 
     
     
         164 . A pharmaceutical composition comprising the nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , or the viral particle of any one of  claims 160 - 162 , and one or more pharmaceutically acceptable excipients. 
     
     
         165 . A method for increasing expression of functional ATP7B protein in a subject comprising administering to said subject the nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , the viral particle of any one of  claims 160 - 162 , or the pharmaceutical composition of  claim 164 . 
     
     
         166 . A method for treating a disorder associated with an ATP7B deficiency comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , the viral particle of any one of  claims 160 - 162 , or the pharmaceutical composition of  claim 164 . 
     
     
         167 . A method for treating Wilson's disease comprising administering to a subject in need thereof a therapeutically effective amount of the nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , the viral particle of any one of  claims 160 - 162 , or the pharmaceutical composition of  claim 164 . 
     
     
         168 . The nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , the viral particle of any one of  claims 160 - 162 , or the pharmaceutical composition of  claim 164  for use in increasing the expression of functional ATP7B in a subject. 
     
     
         169 . The nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , the viral particle of any one of  claims 160 - 162 , or the pharmaceutical composition of  claim 164  for use in increasing treating a disorder associated with an ATP7B deficiency. 
     
     
         170 . The nucleic acid molecule of any one of  claims 134 - 156 , the expression vector of any one of  claims 157 - 159 , the viral particle of any one of  claims 160 - 162 , or the pharmaceutical composition of  claim 164  for use in treating Wilson's disease. 
     
     
         171 . A process of producing a viral particle according to any one of  claims 160 - 162 , comprising: (i) culturing a host cell according to  claim 163  in a culture medium, and (ii) harvesting the viral particles from the cell culture supernatant or the host cells. 
     
     
         172 . The process of  claim 171 , wherein the host cell further comprises (a) a nucleic acid molecule encoding AAV rep and/or cap genes, and/or (b) a nucleic acid molecule comprising viral helper genes. 
     
     
         173 . Use of the nucleic acid molecule of any one of  claims 134 - 156  or the expression vector of any one of  claims 157 - 159  for the production of viral particles.